1. Crop the video for the cage that you will analyze.
  2. Click on FileCapture and from the Desktop upload the video you will analyze.

On the bottom are information necessary for when you do your analysis.

  1. Click on Threshold. On the right-hand side, you will find a control panel for manipulating the imaging of the video. For Threshold Type, click on BinaryInv and choose the value as 85*.* Be sure to target the mouse (shown in white), and everything else should be opaque. You may have to control the threshold levels multiple times if the recording involves a mouse visible through glass, in which light will reflect.

  1. In RoiActivity, select the ROI by making ROI boxes around the tea ball with your computer mouse or touch pad. Afterwards, you can change the shape of the ROI box by clicking on one of the corners.

In order to see where you want to put the boxes, go to Threshold—> Threshold Type: ToZeroInv—> Threshold Value: 255.

This is a photo showing the coordinates of the boxes you made. You are free to change and type in new coordinate numbers as you wish.

  1. Click on Expression Transform. Type in the code: it ? 1 : 0

  2. Start the analysis by clicking Play (on the top left-hand corner). As you see the mouse enter the ROI, in Accumulate, you will see the number of frames when the mouse was in the ROI increase. To be able to see the actual number of accumulated frames (picture below), simply right click on Accumulate, go to "Show Visualizer," and then click on "Bonsai.Design.ObjectTextVisualizer."

***Disclaimer #1: Ideally, the mice will come over to the tea ball to just sniff. However, there are times when mice will: go to the designated ROI box, but not sniff the tea ball, or the mouse's caudal side, or the mouse may simply go on top of the ball and not sniff, and so forth. Accumulate does not automatically account for the actual number of frames when the mouse sniffed; you'll have to count this manually.

One recommended strategy is to manually track the increasing accumulation of frames when sniffing and non-sniffing are involved.

Example: Suppose you were analyzing sniff time from a video with 100 frames total. Suppose the mouse was in the ROI between Frames #1 and 75 had non-sniffing activities. Then the mouse meanders in other parts of the cage. Be sure to mark something like first 75 frames non-sniffing activity. Then, the mouse comes back to the ROI in Frames #76-100, which actually involved sniffing. Be sure to mark this down too. When all is done, simply subtract the 75 frames (non-sniffing) from the 100 total frames and you get 25 frames where sniffing actually occurred.

***Disclaimer #2: You're welcome to hit the pause button from "File Capture." However, be sure not to pause when the mouse is in the ROI, otherwise Accumulate will keep accumulating an increasing number of frames (check wording here).